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王鹏, 张弩, 张旭杰, 张永安. 虹鳟Fc受体FcγR的α和γ亚基基因的克隆及表达分析[J]. 水生生物学报, 2019, 43(1): 27-36. DOI: 10.7541/2019.004
引用本文: 王鹏, 张弩, 张旭杰, 张永安. 虹鳟Fc受体FcγR的α和γ亚基基因的克隆及表达分析[J]. 水生生物学报, 2019, 43(1): 27-36. DOI: 10.7541/2019.004
WANG Peng, ZHANG Nu, ZHANG Xu-Jie, ZHANG Yong-An. MOLECULAR CLONING AND EXPRESSION ANALYSIS OF THE α AND γ SUBUNIT GENES OF FcγR IN RAINBOW TROUT (Oncorhynchus mykiss)[J]. ACTA HYDROBIOLOGICA SINICA, 2019, 43(1): 27-36. DOI: 10.7541/2019.004
Citation: WANG Peng, ZHANG Nu, ZHANG Xu-Jie, ZHANG Yong-An. MOLECULAR CLONING AND EXPRESSION ANALYSIS OF THE α AND γ SUBUNIT GENES OF FcγR IN RAINBOW TROUT (Oncorhynchus mykiss)[J]. ACTA HYDROBIOLOGICA SINICA, 2019, 43(1): 27-36. DOI: 10.7541/2019.004

虹鳟Fc受体FcγR的α和γ亚基基因的克隆及表达分析

MOLECULAR CLONING AND EXPRESSION ANALYSIS OF THE α AND γ SUBUNIT GENES OF FcγR IN RAINBOW TROUT (Oncorhynchus mykiss)

  • 摘要: Fc受体(FcR)是一种表达在免疫细胞表面的受体分子, 由多亚基构成, 通过与免疫球蛋白(Ig)的Fc段结合引起包括炎症因子释放和吞噬作用等体液和细胞免疫反应。研究采用RACE技术首次克隆得到了虹鳟FcγR的α亚基基因(FcγRα)和γ亚基基因(FcRγ)的cDNA序列, 采用生物信息学软件对FcγRαFcRγ的序列进行了特征分析, 实时荧光定量PCR检测了其在不同组织和细胞亚群中以及在Poly (I鲶C)和LPS刺激后头肾中的表达。结果显示:FcγRα的cDNA全长1677 bp, 开放阅读框为954 bp, 编码317个氨基酸; FcγRα由信号肽和2个Ig样结构域构成, 但没有跨膜区和胞内区。FcRγ亚基存在2种形式, 分别命名为FcRγ1和FcRγ2(包含FcRγ2a和FcRγ2b两个剪接异构体), 它们均由信号肽、跨膜区和胞内的免疫受体酪氨酸活化基序(ITAM)构成。氨基酸序列相似性分析表明虹鳟FcγRα与斑点叉尾鮰FcRI相同率最高(30%), 虹鳟FcRγ1和FcRγ2a/2b与哺乳动物FcRγ相同率最高可达40%。组织表达显示FcγRαFcRγ1和FcRγ2a/2b在头肾、脾脏和血液中表达较高; 细胞亚群表达显示FcγRαFcRγ1和FcRγ2a/2b在髓样细胞群中表达最高; LPS和Poly (I鲶C)刺激后,FcγRαFcRγ1和FcRγ2a/2b在头肾中的表达显著上调, 这表明FcγR在机体抗细菌和抗病毒免疫中可能发挥重要作用。

     

    Abstract: In this study, the cDNA sequences of FcγRα and FcRγ subunits of FcγR rainbow trout were cloned for the first time by RACE technique. The sequences of FcγRα and FcRγ were analyzed by using bioinformatics software. The expressions of the genes in different cell subpopulations and tissues, as well as in the head kidney after Poly (I鲶C) and lipopolysaccharide (LPS) stimulations were analyzed by real-time fluorescent quantitative polymerase chain reaction (PCR). The results showed that the full-length cDNA ofFcγRα is 1677 bp with an open reading frame (ORF) of 954 bp encoding 317 amino acids. The FcγRα is composed of a signal peptide and two Ig-like domains, however, without including transmembrane and intracellular regions. There are three kinds of FcγR subunit, involving FcRγ1, FcRγ2a, and FcRγ2b. FcRγ1 and FcRγ2 genes are located on different chromosomes, while FcRγ2a and FcRγ2b are two splicing isoforms of FcRγ2 gene. These subunits are composed of signal peptide, transmembrane domain, and immunoreceptor tyrosine-based activation motif (ITAM). Amino acid sequence similarity analysis showed that the highest identity (30%) of FcγRα is between rainbow trout and channel catfish (Ictalurus punctatus), and FcRγ1 and FcRγ2a/2b in rainbow trout have the maximum sequence identity (40%) with mammalian FcRγ. Tissue distribution analysis showed that the expression of FcγRα, FcRγ1, and FcRγ2a/2b was higher in head kidney, spleen, and blood cells, respectively. Analysis of cell subpopulations showed that the expression of FcγRα, FcRγ1, and FcRγ2a/2b was the highest in the myeloid cell population. In addition, the expression of FcγRα, FcRγ1, and FcRγ2a/2b in the head kidney was significantly up-regulated after LPS and Poly (I鲶C) stimulations, indicating that FcγR plays an important role in the antibacterial and antiviral immunity.

     

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